Synthesis of disaccharide fragments of the AT-III binding domain of heparin and their sulfonatomethyl analogues

Article abstract of DOI:10.1016/j.carres.2011.06.021

D-Glucuronate and L-iduronate-containing disaccharides related to the antithrombin-binding domain of heparin were prepared. The carboxylic function of the uronic acid unit was formed on a disaccharide level in the case of the glucuronate, while on a monos

Full text of DOI:10.1016/j.carres.2011.06.021

Carbohydrate Research 346 (2011) 1827–1836
Contents lists available at ScienceDirect
Carbohydrate Research
journal homepage: www.elsevier.com/locate/carres
Synthesis of disaccharide fragments of the AT-III binding domain of heparin
and their sulfonatomethyl analogues
Mihály Herczeg a, László Lázár a, Attila Mándi a,b, Anikó Borbás a, , István Komáromi c, András Lipták a,
⇑
Sándor Antus a,b
a Research Group for Carbohydrates of The Hungarian Academy of Sciences, University of Debrecen, PO Box 94, H-4010 Debrecen, Hungary
b Department of Organic Chemistry, University of Debrecen, PO Box 20, H-4010 Debrecen, Hungary
c Thrombosis, Haemostasis and Vascular Biology Research Group of The Hungarian Academy of Sciences, University of Debrecen, H-4032 Debrecen, Hungary
a r t i c l e i n f o
a b s t r a c t
Article history:
D-Glucuronate and L-iduronate-containing disaccharides related to the antithrombin-binding domain of
Received 10 November 2010
Received in revised form 8 June 2011
Accepted 20 June 2011
heparin were prepared. The carboxylic function of the uronic acid unit was formed on a disaccharide level
in the case of the glucuronate, while on a monosaccharide level in the case of the iduronate derivatives.
Synthesis of their sulfonic acid analogues was carried out analoguosly applying sulfonatomethyl-
containing acceptors in the form of either salts or methyl esters. Significant difference could be observed
in the methyl ether formation reactions of the sulfonatomethyl-containing uronate disaccharides and the
non-sulfonic acid uronates.
Available online 24 June 2011
Keywords:
Heparin
Heparinoid disaccharides
Ó 2011 Elsevier Ltd. All rights reserved.
D-Glucuronic acid
L
-Iduronic acid
Sulfonatomethyl analogues
1. Introduction
simplified structures much easier to synthesize than fondaparinux.
In addition, idraparinux5,6 (3a), the most potent representative of
Heparin, a highly sulfated linear polysaccharide belonging to
the family of glycosaminoglycans is a well-known blood anticoag-
ulant employed extensively in medical practice.1 The anticoagulant
action of heparin arises from its ability to accelerate the inhibitory
activity of antithrombin III (AT-III), a serine protease inhibitor that
blocks thrombin and factor Xa in the blood-coagulation cascade.2
After identification of the exact sequence of heparin that associates
with the AT-III (1) a concerted drug development programme has
been undertaken based on extensive structure–activity studies
using synthetic oligosaccharides.3 These efforts resulted in fonda-
parinux (2), a synthetic pentasaccharide heparin analogue which
is used as a new antithrombotic drug under the name Arixtra.4
Fondaparinux selectively inhibits factor Xa, and it has longer
half-life and higher anti-Xa activity than heparin and low molecu-
these analogues possesses increased anticoagulant activity and
longer duration of action (Fig. 1).4a,5 It turned out, however, that
the extremely long elimination half-life of idraparinux (ꢀ60 days
by clinical trials) may lead to serious bleeding complications,
therefore, its development was terminated.7 Idrabiotaparinux
(3b), the biotinylated form of 3a has been developed with the
aim to improve the management of bleeding events seen with idra-
parinux.8 Idrabiotaparinux displays the same advantageous prop-
erties as idraparinux and has the added safety feature of easy
and rapid neutralization of its anticoagulant effect by addition of
the specific antidote, avidin.7,8
Our research group decided the synthesis of the isosteric sul-
fonic acid analogues of idraparinux (2) by systematic replacement
of the sulfate esters with sulfonatomethyl moieties to obtain new
selective Xa inhibitors and to acquire further information on the
structure–activity relationship of the antithrombotic action of hep-
arin pentasaccharide. It is known that the anionic groups are
essential for the activation of AT-III, and that the type of charge
is also crucial,3c however, replacement of the sulfate group with
sulfonic acid moiety has not been investigated till now.
lar-weight heparin. However, its synthesis represents
challenge, due to the presence of -glucuronic acid, -iduronic acid
and -glucosamine, most of them N- and O-sulfated on selected
a real
D
L
a-D
positions. Based on the observations of SAR studies that the
N-sulfate groups could be replaced by O-sulfates, and the free
hydroxyl groups could be masked with alkyl groups, a family of
non-glycosaminoglycan derivatives was created,5 having
Synthesis of sulfonatomethyl analogues of the EF and GH
disaccharides (5–7 and 9–11)9 as well as sulfonatomethyl
analogues10 of the DEF trisaccharide of compound 3a have been
reported recently. Here, we present the synthesis of two uronic
acid-containing fragments of idraparinux (4 and 8) together with
⇑
Corresponding author. Tel.: +36 52512900/22257; fax: +36 52512900/22342.
E-mail addresses: borbasa@puma.unideb.hu, borbas.aniko@science.unideb.hu
(A. Borbás).
0008-6215/$ - see front matter Ó 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.carres.2011.06.021

1828
M. Herczeg et al. / Carbohydrate Research 346 (2011) 1827–1836
OBn
O
OBn
O
D
E
F
G
H
R2
OAc
O
a
R1O
R1O
-
O
-
-
OSO3
O
O
AcO
AcO
HO
+
OSO3
O
OSO3
BnO
-OOC
O
BnO
COO-
OH
O
O
OR1
BnO
O
BnO
AcO
O
OMe
OMe
Br
HO
HO
O
O
12
-O3SO
13
1
14 R1 = Ac, R2 = CH2OAc
R2HN
OR
b
c
HO
HO
-O SHN
-O3SO
3
1 = H, R2 = CH2OH
1 =H , R2 = COO-
-O3SHN
OH
15
16
R
R
1 R1 = H, R2 = Ac
COO-
OR
COO-
O
OBn
O
2 R1 = Me, R2 = SO3
-
O
d
O
MeO
MeO
O
O
+
RO
BnO
MeO
-
-
RO
-
OMe
OSO3
O
OSO3
BnO
OMe
OSO3
OMe
OMe
COO-
17 R = Bn
-OOC
O
18
e
OMe
O
O
O
O
19 R = H
O
MeO
MeO
O
O
-O3SO
R
-O3SO
-O SO
3
MeO
f
OMe
MeO
-O3SO
MeO
4
3a R = OMe
O
S
Scheme 1. Synthesis of the EF fragment of idraparinux by selective oxidation.
Reagents and conditions: (a) CH2Cl2–toluene, 4 Å Ms, AgOTf, ꢂ30 °C to rt, 24 h
(94%); (b) MeOH, NaOMe, rt, 2 h (97%); (c) satd NaHCO3, KBr, NaOCl, TEMPO, 0 °C to
rt, 2 h (78%); (d) DMF, NaH, MeI, 0 °C to rt, 2 h (17: 42%, 18: 40%); (e) ethanol, Pd(C),
H2, 10 atm, rt, 12 h (94%); (f) DMF, SO3ꢁpy, 0 °C to rt, 5 h (70%).
3b R =
NH
HN
NH
O
O
NH
Figure 1. Structures of the antithrombin-binding domain of heparin (1), synthetic
antithrombotic drug fondaparinux (2), the non-glycosaminoglycan analogue idra-
parinux (3a) and its biotinylated derivative idrabiotaparinux (3b).
In order to avoid the elimination side reaction caused by meth-
ylation of the base-sensitive uronate derivative, compound 17 was
prepared from 15 by permuting the oxidation and methylation
steps. The primary hydroxyl of 15 was protected temporarily in
form of trityl ether, then the secondary hydroxyls of 20 were meth-
ylated to afford 21. Removal of the trityl group gave 22 whose
TEMPO-based oxidation resulted in the fully protected glucuronide
17 in good yield (Scheme 2). Synthesis of 4 starting from 12 and 13
through the 6-O-trityl intermediate 20 required eight steps and re-
sulted in the target glucuronide disaccharide with 27% overall
yield.
To obtain the sulfonic acid analogues of disaccharide 4, first the
sulfonatomethyl-containing glucoside acceptors have to be pre-
pared.9 The key step of their synthesis was the addition of hydro-
gen sulfite to the exomethylene moiety of the appropriate
glucoside resulting exclusively in the primary alkane sulfonate in
a radical chain reaction using tert-butyl peroxybenzoate as radical
initiator.14,15
Previously, the 2-deoxy-2-sulfonatomethyl and the 3-deoxy-3-
sulfonatomethyl acceptors (24 and 27) have been prepared by
the following sequence of reactions: the exomethylene derivatives
2316 and 26,17 were reacted with NaHSO3, respectively, followed
by regioselective hydrogenolysis of the 4,6-O-benzylidene acetal.
However, the 4-OH derivatives 24 and 27 could be isolated only
with moderate yields due to partial hydrolysis of the benzylidene
acetal under the slightly acidic conditions of the first addition
step.9 We attempted to increase the yield of the two-step proce-
dure by reversing the acetal reduction and the addition. In the case
of the 3-exomethylene derivative 26 regioselective opening of the
4,6-O-acetal ring using a triethylsilane–BF3ꢁEt2O reagent combina-
tion18 (28), and subsequent sulfonation afforded the desired 27
with increased yield compared to the previous pathway (65% vs
47% over two steps). However, treatment of 23 with Et3SiH/
BF3ꢁEt2O resulted in the unexpected 2-methyl glycal 25 as the main
unpublished details of the synthesis of their sulfonic acid mimetics
(Fig. 2). Differences in the synthesis of disaccharide uronates with
or without sulfonic acid content are also discussed.
2. Results and discussion
For the synthesis of compound 4 the b-(1?4)-linked skeleton
was prepared by coupling of donor 1211 and acceptor 1312 and
the disaccharide 14 was isolated in high yield. Zemplén deacetyla-
tion of 14 afforded the tetrahydroxy derivative 15. Selective
oxidation of the primary hydroxyl group of compound 15 by
TEMPO-based oxidation method13 using sodium-hypochlorite as
co-oxidant gave compound 16. Treatment of 16 with methyl iodide
and sodium hydride to achieve methylation of the three hydroxyl
groups of the uronic acid residue resulted in a 3:2 mixture of the
desired product 17 and the by-product 18 formed by b-elimina-
tion. Catalytic hydrogenation of 17 afforded the triol 19 which
was O-sulfated to obtain the target disaccharide 4 (Scheme 1).
E
F
G
H
OSO3
-
-
OSO3
O
COO-
O
O
O
O -O3SO
-O3SO
OMe
O
MeO
MeO
-O3SO
-O3SO
-OOC
OMe
OMe
OMe
4
OMe OMe
8
-
OSO3
-
COO-
OSO3
O
O
-O3SO
O
O
O
MeO
MeO
MeO
-O3SO
-OOC
-O3S
O
OMe
MeO
-O3S
OMe
MeO MeO
5
9
-
OSO3
O
-
COO-
OSO3
O
O
O
O
-O3S
MeO
MeO
MeO
OR2
O
OBn
O
-O3SO
-OOC
OH
O
OBn
O
O
-O3SO
-
MeO
HO
HO
R1O
R1O
a
d
SO3
10
OMe
O
O
OMe
-
MeO MeO
17
BnO
BnO
6
-
OR1
SO3
BnO
BnO
OH
OMe
OMe
SO3
1
15
R
= H, R2 = CPh3
COO-
O
20
O
b
c
-O3SO
-O3SO
MeO
-OOC
21 R1 = Me, R2 = CPh3
22 R1 = Me, R2 = H
O
O
O
MeO
MeO
-O3SO
O
-O3SO
OMe
MeO
OMe
MeO MeO
7
11
Scheme 2. Improved synthesis of the methylated uronate 17. Reagents and
conditions: (a) py, TrCl, 0 °C to rt, 24 h (83%); (b) DMF, NaH, MeI, 0 °C to rt, 2 h
(86%); (c) 80% AcOH, 50 °C, 2 h (98%); (d) satd NaHCO3, KBr, NaOCl, TEMPO, 0 °C to
rt, 2 h (64%).
Figure 2. Structures of the EF and the GH fragments of idraparinux (4 and 8) and
their isosteric sulfonic acid derivatives (5–7 and 9–11).

M. Herczeg et al. / Carbohydrate Research 346 (2011) 1827–1836
1829
product. The reaction can be explained by the formation of a stabi-
lized allylic cation (allyl oxocarbenium ion) whose reaction with
the hydride donor reagent gave the 2-deoxy-2-methyl glycal deriv-
ative (Scheme 3).
Cleavage of the glycosidic bond upon reductive opening of
benzylidene-type acetals is extremely rare, however, not
unprecedented.19,20
The 2-(sodium sulfonatomethyl)- and 3-(sodium sulfonatom-
ethyl)-containing analogues of disaccharide 4 have been synthe-
sized through the appropriate 60-O-trityl intermediates, starting
from the acetobromo glucose 12 and acceptors 24 and 27, respec-
tively, as we published recently.9 It is worth to mention that the
sulfonic acid salts proved to be excellent acceptors, and similar
overall yields could be achieved for the sulfonic acid-containing
disaccharides 5 and 6 as for compound 4.
However, preparation of the 6-(sodium sulfonatomethyl)-con-
taining derivatives 7 via tritylation was failed. Glycosylation of
the sulfonic acid heptoside 299 in a salt form with the acetobromo
glucose donor 12 upon silver triflate activation and the following
deacetylation took place smoothly affording the desired disaccha-
ride 30 in a 48% yield.9 However, subsequent tritylation of 30
Figure 3. Superposition of the six lowest energy conformations of 30 from the high
temperature molecular dynamics (ball-and-stick representation: lowest energy
conformer; stick representation: conformers 2–6).
failed, despite
a
high excess of the reagent, addition of
30
dimethylamino pyridine and prolongation of the reaction time
(Scheme 4). Silylation using tert-butyldiphenylsilyl chloride was
also unsuccessful.
Geometry optimization on the structures sampling from high
temperature molecular dynamics simulations trajectory for 30
gave explanation for the failure of substitution of the 60-hydroxyl
group. In the lowest energy conformations the hydroxyl group is
inaccessible by the bulky reagents being in a pocket formed by
a
-
SO3
COO-
O
O
O
RO
RO
BnO
BnO
OR
OMe
31 R = H
b
32 R = Me
c, d
7
OBn
O
O
Ph
Ph
O
O
Scheme 5. Synthesis of the 6-deoxy-6-sulfonatomethyl analogue of the EF
fragment of idraparinux by selective oxidation. Reagents and conditions: (a) satd
NaHCO3, KBr, NaOCl, TEMPO, 0 °C to rt, 2 h (91%); (b) DMF, NaH, MeI, 0 °C to rt, 2 h
(79%); (c) ethanol, Pd(C), H2, 10 atm, rt, 12 h (96%); (d) DMF, SO3ꢁpy, 0 °C to rt, 5 h
(65%).
b
O
a, b
O
HO
BnO
O
BnO
BnO
H2C
-O3S
OMe
OMe
CH3
25
23
24
OBn
O
OBn
O
O
Ph
O
a
O
b
a, b
HO
HO
27
H2C
BnO
the two benzyl groups and the sugar ring (Fig. 3). These conforma-
tions are probably stabilized by hydrogen bonds between the sul-
fonate moiety and the 20- or 30-hydroxyls. (For the most stable
geometry the corresponding distances between the sulfonate oxy-
gens and the 20- or 30-hydroxyl hydrogens are 1.84 Å for 20-OH and
1.91 Å for 30-OH from DFT calculations while 1.77 Å for 20-OH and
1.64 Å for 30-OH were predicted by the AMBER-GAFF force field.)
The above assumptions were confirmed by NMR experiments:
the hydrogen of the 60-OH could be exchanged for deuterium
immediately, while deuteration of 20-OH and 30-OH was slow.
Since selective protection of the sterically hindered 60-hydroxyl
group failed, the tetrahydroxy derivative 30 was converted directly
into the uronic acid 31 by TEMPO-based selective oxidation. Meth-
ylation of 31 with methyl iodide and sodium hydride was consid-
ered to be a risky step that might lead to b-elimination of the
uronic acid residue as it happened to 16. Surprisingly, methylation
in the presence of the strong base proceeded smoothly and no
elimination was observed (Scheme 5). The obtained 32 could be
converted into the targeted 6-deoxy-6-sulfonatomethyl-contain-
ing disaccharide 7 by catalytic hydrogenation and subsequent sul-
fation of the liberated two hydroxyls. Applying the selective
oxidation method a 22% overall yield could be obtained for 7 in a
six-step procedure.9
BnO
H2C
-O3S
BnO
OMe
OMe
OMe
27
26
28
Scheme 3. Previously described (a followed by b) and recent (b followed by a)
reaction routes for the 2-deoxy-2-sulfonatomethyl and the 3-deoxy-3-sulfonatom-
ethyl acceptors. Reagents and conditions: (a) 70% EtOH, tBu–peroxybenzoate,
NaHSO3, reflux, 4 h (55% from 23, 53% from 26, 74% from 28); (b) CH2Cl2, Et3SiH,
BF3ꢁEt2O, 0 °C to rt, 4 h (24: 87%, 27: 88%, 25: 41%, 28: 78%).
-O3S
O
HO
12 +
BnO
BnO
OMe
29
a, b
-
SO3
OH
O
HO
HO
O
O
BnO
OH
BnO
OMe
30
c, d
For the incorporation of uronic acid residues into oligosaccha-
rides two distinct strategies exist: glycosylation followed by oxida-
tion (post-glycosidation oxidation) and oxidation of the
monosaccharide building blocks followed by glycosylation
Scheme 4. Attempted synthesis of the 6-deoxy-6-sulfonatomethyl analogue of the
EF fragment of idraparinux via tritylation. Reagents and conditions: (a) CH2Cl2–
toluene, 4 Å Ms, AgOTf, ꢂ30 °C to rt, 24 h (64%); (b) MeOH, NaOMe, rt, 2 h (75%); (c)
TrCl (3 equiv), py, DMAP; (d) TBDPSCl, imidazole, DMF.

1830
M. Herczeg et al. / Carbohydrate Research 346 (2011) 1827–1836
(pre-glycosidation oxidation).21 Although the glucuronide-contain-
ing disaccharides were prepared successfully by post-glycosidation
oxidation, the other strategy that is glycosylation with an iduronic
donor was selected to synthesize our targeted GH analogue disac-
charides 8–11. Since neither idose nor iduronic acid is readily
available from natural and commercial sources and preparation
of both type of donors requires a lengthy synthetic procedure,22
the pre-glycosidation oxidation seemed to be more efficient.
+
39
13
a
OBn
O
O
OMe
O
BnO
MeOOC
OR
BnO
OMe
OR
40 R = Ac
b
c
41
R = H
42 R = Me
L
-Iduronic acid trichloroacetimidate 39 was prepared on the ba-
sis of the well-established procedure elaborated for the appropri-
ate 3-O-benzyl-
-iduronate donor.22h,23 The iduronic acid 36 was
prepared from the corresponding -glucofuranuronic acid deriva-
d
OR
O
L
O
D
OMe
O
RO
tive24 by means of a two-step epimerization at C-5.23 Deisopropyli-
denation of 36 with trifluoroacetic acid afforded the desired
hemiacetal as a mixture of pyranoses and furanoses. Acetylation
of the crude reaction mixture, followed by column chromatogra-
phy gave 37 with 36% yield over the two steps. Selective deacety-
lation of the anomeric position (38) and imidate formation then
resulted in the iduronic acid donor 39 (Scheme 6).
-OOC
RO
OMe
OMe OMe
43
R = Bn
e
44 R = H
f
8
Coupling of the acceptor 13 with the imidate donor 39 upon tri-
methylsilyl triflate activation afforded the disaccharide 40 and its
Zemplén deacetylation resulted in diol 41. Attempted methylation
with methyl iodide and sodium hydride failed leading to complete
b-elimination of the iduronic acid residue, therefore, the strong
base was exchanged into freshly prepared silver(I)oxide. Under
such conditions, the methylated 42 was obtained in high yield
and no elimination occurred. The uronic ester was treated with
aqueous sodium hydroxide to give the sodium uronate 43. Re-
moval of the benzyl protecting groups by catalytic hydrogenation
resulted in the triol 44 which upon sulfation with the SO3ꢁpyridine
complex furnished the GH fragment of idraparinux (8, Scheme 7).
Overall yield for 8 was 37% starting from 39 and 13.
Coupling of the sulfonatomethyl glucosides in salt form (24, 27
and 29) with the iduronic acid donor failed, therefore, the sulfonic
acids in the form of the methyl esters were resorted as the accep-
tors. The methyl ester 45 was prepared from the salt 24 by libera-
tion of the sulfonic acid followed by esterification with ethereal
diazomethane.9 Glycosylation of the 2-deoxy-2-sulfonatomethyl
Scheme 7. Synthesis of the GH fragment of idraparinux. Reagents and conditions:
(a) CH2Cl2, 4 Å Ms, TMSOTf, ꢂ20 °C, 30 min (94%); (b) MeOH, NaOMe, rt, 1 h (95%);
(c) DMF, Ag2O, MeI, rt, 96 h (76%); (d) 0.1 M NaOH, rt, 24 h (68%); (e) ethanol, Pd(C),
H2, rt, 12 h (85%); (f) DMF, SO3ꢁpy, 0 °C to rt, 5 h (94%).
carboxylic ester giving sodium uronate. Catalytic hydrogenation
and subsequent sulfation afforded the 2-deoxy-2-sulfonatometh-
yl-containing disaccharide 9 with 55% overall yield over eight steps
(Scheme 8).9
The synthesis of the 3-sulfonatomethyl- and the 6-sulfonatom-
ethyl-containing iduronides 10 and 11 has been published in de-
tail.9 Their preparation was carried out analogously to compound
9, except for the the methylation step. It is important to note that
methylation of diols 50 and 51 using sodium hydride and methyl
iodide did not cause elimination side reaction and afforded exclu-
sively the desired products 52 and 53, respectively (Scheme 9). In
the end 42% overall yield could be achieved for 10 and 48% for 11
starting from the iduronic acid donor and the methyl sulfonatom-
ethyl-containing acceptors.
glucoside acceptor 45 with the donor 39 afforded the desired
a
-
In conclusion, the EF and GH disaccharide fragments of
idraparinux were prepared following two different strategies. The
glucuronic acid containing EF disaccharide was synthesized by
post-glycosidation oxidation using an acetobromo glucose donor
to build up the disaccharide skeleton and TEMPO-based oxidation
to form the carboxylic function at a disaccharide level. Preparation
of the iduronic acid-containing GH disaccharide was carried out by
pre-glycosidation oxidation using an iduronic acid donor to
achieve the disaccharide uronate.
Synthesis of the sulfonic acid analogues could be carried out
analoguosly applying sulfonatomethyl-containing acceptors in
the form of salts and methyl esters. The sulfonatomethyl gluco-
sides in the form of salts were efficient acceptors when reacting
with the acetobromo glucose donor. The glycosylation reactions
failed when the sulfonic acid salt acceptors were reacted with
the iduronic acid donor with inherently low reactivity. In that case
the sulfonic acid acceptors in the form of esters were applied to
effectuate the glycosylations in high yields.
linked disaccharide whose deacetylation gave 46 in a high yield.
Usual methylation was attempted applying sodium hydride and
methyl iodide, since similar methylation of the sulfonic acid-con-
taining glucuronate 31 took place smoothly. However, with 46 an
inseparable mixture of the desired product 47 and the by-product
48 (as a result of b-elimination of the base-sensitive iduronic acid
residue) was formed in a ratio of ꢀ3:1. To overcome this difficulty,
methylation of 46 was carried out under slightly acidic conditions
with diazomethane25 applying silica gel25b or BF3ꢁEt2O25c to cata-
lyse the reaction. The silica gel promoted methylation proved to
be more efficient affording the desired product 4949 in high yield.
Compound 49, carrying two ester functions, was converted into
the disodium salt in two steps; the sulfonic ester was treated with
sodium iodide in acetone to give sodium sulfonate in a nucleophilic
substitution reaction, then the crude product was reacted
with aqueous sodium hydroxide in methanol to hydrolyse the
Comparing the efficacy of the synthesis of disaccharides with or
without sulfonic acid content similar overall yields could be
reached for the non-sulfonic acid disaccharides (4 and 8) and their
sulfonatomethyl-containing analogues (5–7 and 9–11).
Interestingly, the sulfonatomethyl-containing uronate disac-
charides proved to be significantly more stable under the basic
conditions of methylation than the non-sulfonic acid uronates.
There was only one precedent for b-elimination of the base-sensi-
tive uronates of the sulfonic acid analogues (namely, the reaction
of 46 with sodium hydride and methyl iodide). In that case,
COOMe
OMe
O
OMe
O
OR1
O
OH
CCl3
MeOOC
MeOOC
O
a,b
d
OMe
NH
OR2 OR2
OAc OAc
O
O
37 R1 = R2 = Ac (α:β~1:10)
39
c
36
1
38
R
=H, R2 = Ac
Scheme 6. Synthesis of the
F3CCOOH, rt, 15 min; (b) py, Ac2O, 0 °C to rt, 24 h (36% for two steps); (c) THF,
BnNH2, rt, 3 h (79%); (d) CH2Cl2, DBU, trichloroacetonitrile, 0 °C, 30 min (72%).
L
-iduronate donor. Reagents and conditions: (a) 90%

M. Herczeg et al. / Carbohydrate Research 346 (2011) 1827–1836
1831
OBn
OBn
O
OBn
O
O
O
a,b
O
d
MeO
HO
BnO
BnO
OMe
O
BnO
O
MeOOC
MeOOC
OMe
OMe
MeO3S
MeO3S
OMe
MeO3S
HO
OH
46
OMe OMe
49
45
c
e-h
OBn
O
OBn
O
O
O
BnO
MeOOC
MeO
BnO
-O3S
OMe
O
9
+
O
MeOOC
-O3S
OMe
OMe
MeO MeO
OMe
48
47
Scheme 8. Synthesis of the 2-deoxy-2-sulfonatomethyl analogue of the GH fragment of idraparinux. Reagents and conditions: (a) 39, CH2Cl2, 4 Å Ms, TMSOTf, ꢂ20 to 0 °C,
15 min (80%); (b) MeOH, NaOMe, rt, 2 h (97%); (c) DMF, NaH, MeI (d) CH2Cl2, CH2N2, silicagel, 0 °C (88%); (e) acetone, NaI, rt, 2 h; (f) 0.1 M NaOH, rt, 24 h (96% over two steps);
(g) ethanol, Pd(C), H2, 10 atm, rt, 12 h (97%); (h) DMF, SO3ꢁpy, 0 °C to rt, 5 h (87%).
optimizations using the suitably developed GAFF empirical force
field on the equidistantly saved 100,000 trajectory snapshot geom-
etries were carried out by means of the AMBER molecular dynam-
ics simulation package.26,20 The B3LYP/6-31G(d) density functional
calculation on the lowest energy conformer was carried out using
the GAUSSIAN 03 package.27 Ball-and-stick and stick representations
of the conformers were generated by the VMD software.28
OBn
O
SO3Me
O
O
MeO
O
MeO
BnO
O
BnO
SO3Me
MeOOC
OMe
BnO
O
MeOOC
OMe
OH OH
OH OH
50
51
NaH
MeI
DMF
NaH
MeI
DMF
3.2. Methyl (2,3,4,6-tetra-O-acetyl-b-D-glucopyranosyl)-(1?4)-
SO3Na
O
OBn
O
2,3,6-tri-O-benzyl- -glucopyranoside (14)
a-D
O
MeO
O
MeO
BnO
O
BnO
SO3Na
OMe OMe
To a mixture of compound 13 (1.00 g, 2.16 mmol) and acetob-
romoglucose (12, (1.78 g, 4.32 mmol)) in dry CH2Cl2 (20 mL) pow-
dered 4 Å molecular sieves (1.5 g) was added. The stirred mixture
was cooled to 0 °C under argon. After 1 h at this temperature,
AgOTf (1.2 g, 4.68 mmol) dissolved in toluene (4 mL) was added.
After another hour the reaction was allowed to warm up to room
temperature and stirred overnight. The reaction mixture was fil-
tered through a pad of Celite. After filtration Et3N (1 mL) was
added, and the solvents were evaporated in vacuo. The crude prod-
uct was purified by silica gel chromatography (1:1 n-hexane–
MeOOC
BnO
OMe
O
MeOOC
OMe OMe
53 (88%)
OMe
52 (83%)
Scheme 9. Synthesis of the 3-deoxy-3-sulfonatomethyl and 6-deoxy-6-sulfon-
atomethyl analogues of compound 8.
methylation was carried out under acidic conditions applying ethe-
real diazomethane in the presence of silica gel.
Synthesis of the pentasaccharide analogues of idraparinux by
applying the presented methods, as well as biological investiga-
tions of the prepared sulfonic acid derivatives is in progress.
EtOAc) to give 14 (1.61 g, 94%) as a colourless syrup. [
a
]
ꢂ6.2 (c
D
0.11, CHCl3); Rf 0.42 (1:1 n-hexane–EtOAc); 1H NMR (CDCl3,
360 MHz): d (ppm) 7.38–7.27 (m, 15H, arom.), 5.01–4.39 (m,
10H), 4.15 (m, 1H), 3.91–3.35 (m, 9H), 3.37 (s, 3H, OCH3), 2.00,
1.98, 1.95 (3 ꢃ s, 12H, 4 ꢃ CH3); 13C NMR (CDCl3, 90 MHz): d
(ppm) 170.4, 170.0, 169.2, 168.8 (4C, 4 ꢃ CO), 139.2, 138.1, 137.5
(3C, 3 ꢃ Cq arom.), 128.5–127.0 (arom.), 99.8, 98.2 (C-1, C-10),
79.7, 78.7, 77.1, 73.0, 71.7, 71.3, 69.5, 67.9 (skeleton carbons),
74.9, 73.5, 73.3 (3 ꢃ PhCH2), 67.4 (C-60), 61.5 (C-6), 55.2 (OCH3),
20.5, 20.4, 20.3 (4 ꢃ CH3); MALDI-TOF (positive ion): m/z 817.17
[M+Na]+ (calcd 817.30). Anal. Calcd for C42H50O15 (794.84): C,
63.47; H, 6.34. Found: C, 63.33; H, 6.29.
3. Experimental
3.1. General information
Optical rotations were measured at room temperature with a
Perkin-Elmer 241 automatic polarimeter. TLC was performed on
Kieselgel 60 F254 (Merck) with detection by immersing into 5% eth-
anolic sulfuric acid soln followed by heating. Column chromatogra-
phy was performed on Silica Gel 60 (Merck 0.063–0.200 mm) and
Sephadex LH-20 (Sigma-Aldrich, Bead size 25–100
l). Organic
3.3. Methyl (b-
D-glucopyranosyl)-(1?4)-2,3,6-tri-O-benzyl-a-D-
solutions were dried over MgSO4, and concentrated in vacuum.
The 1H (200, 360, 400 and 500 MHz) and 13C NMR (50.3, 90.54,
100.28, 125.76 MHz) spectra were recorded with Bruker AC-200,
Bruker DRX-360, Bruker DRX-400 and Bruker DRX-500 spectrome-
ters. Chemical shifts are referenced to Me4Si (0.00 ppm for 1H) or to
the residual solvent signals (CDCl3: 77.00 ppm for 13C).
glucopyranoside (15)
To a solution of 14 (1.45 g, 1.82 mmol) in MeOH (25 mL) a cat-
alytic amount of NaOMe (11 mg, 0.2 mmol) was added. After 2 h
stirring, the mixture was neutralized with Amberlite IR-120 H+
ion-exchange resin, filtered and concentrated to give 15 (1.11 g,
MALDI-TOF MS analyses of the compounds were carried out in
the positive reflectron mode using a BIFLEX III mass spectrometer
(Bruker, Germany) equipped with delayed-ion extraction. The ma-
trix solution was a satd 2,4,6-trihydroxy-acetophenone (THAP)
solution in MeCN.
97%) as a colourless syrup. [a] +5.4 (c 0.19, MeOH); Rf 0.42 (9:1
D
CH2Cl2–MeOH); 1H NMR (CD3OD, 360 MHz): d (ppm) 7.44–7.26
(m, 15H, arom.), 4.96 (d, 1H, J 10.3 Hz, PhCH2), 4.76 (d, 1H, J1,2
0
0
3.6 Hz, H-1), 4.70–4.51 (m, 5H, PhCH2), 4.41 (d, 1H, J1 ,2 7.3 Hz,
H-10), 4.00–3.95 (m, 2H), 3.86–3.69 (m, 4H), 3.55–3.46 (m, 2H),
3.37 (s, 3H, OCH3), 3.30–3.17 (m, 4H); 13C NMR (CD3OD,
90 MHz): d (ppm) 139.5 (3 ꢃ Cq arom.), 129.9–128.7 (arom.),
The molecular dynamics simulations (100 ns, 1200 K constant
temperature, 1fs time step) and the preliminary geometry

1832
M. Herczeg et al. / Carbohydrate Research 346 (2011) 1827–1836
103.5 (C-10), 99.0 (C-1), 81.6, 80.6, 78.8, 77.9, 76.5, 75.7, 72.1, 71.6
(skeleton carbons), 77.3, 74.2, 74.1 (3 ꢃ PhCH2), 69.2 (C-6), 63.3 (C-
60), 55.6 (OCH3); MALDI-TOF (positive ion): m/z 649.22 [M+Na]+
(calcd 649.26). Anal. Calcd for C34H42O11 (626.69): C, 65.16; H,
6.76. Found: C, 64.87; H, 6.70.
(calcd 695.24). Anal. Calcd for C36H41NaO11 (672.69): C, 64.28; H,
6.14. Found: C, 64.08; H, 6.09.
3.6. Methyl (sodium 2,3,4-tri-O-methyl-b-D-glucopyranosyluro
nate)-(1?4)- -glucopyranoside (19)
a-D
A mixture of compound 17 (196 mg, 0.287 mmol) and Pd/C
(10%, 80 mg) in 96% EtOH–AcOH (19:1, 15 mL) was stirred in an
autoclave under H2 atmosphere (at 10 bar) for 12 h. The catalyst
was filtered off through a pad of Celite and the filtrate was concen-
trated under reduced pressure to give 19 (111 mg, 94%) as a white
3.4. Methyl (sodium b-
D-glucopyranosyluronate)-(1?4)-2,3,6-
tri-O-benzyl- -glucopyranoside (16)
a
-D
A stirred mixture of compound 15 (200 mg, 0.319 mmol) satd
NaHCO3 solution (2.6 mL), KBr (20 mg, 0.16 mmol) and TEMPO
(5 mg, 0.031 mmol) was cooled to 0 °C, and then 5% NaOCl solution
(2.6 mL, ꢀ1.75 mmol) was added slowly in portions. After 2 h the
reaction mixture was extracted with CH2Cl2 (10 mL), and the aque-
ous phase was concentrated. Then MeOH (10 mL) was added and
after stirring for 20 min, the insolubles were removed by filtration.
The filtrate was concentrated under reduced pressure. The crude
product was purified by silica gel chromatography in 85:15
CH2Cl2–MeOH, to give the uronic acid salt 16 as a white powder
powder. [
a]
D +58.6 (c 0.11, MeOH); Rf 0.24 (1:1 CH2Cl2–MeOH); 1H
NMR (CD3OD, 360 MHz): d (ppm) 4.73 (d, 1H, J1,2 3.4 Hz, H-1), 4.50
(d, 1H, J1 ,2 7.1 Hz, H-10), 3.93–3.68 (m, 5H), 3.61, 3.55, 3.52, 3.43
(4 ꢃ s, 12H, 4 ꢃ OCH3), 3.63–3.35 (m, 2H), 3.25–3.07 (m, 3H); 13C
NMR (CD3OD, 90 MHz): d (ppm) 176.6 (CO), 104.2 (C-10), 100.9
(C-1), 87.2, 84.3, 82.7, 80.6, 76.7, 73.3, 73.1, 72.3 (skeleton car-
bons), 61.2, 61.1, 60.8, 55.7 (4 ꢃ OCH3), 61.0 (C-6); MALDI-TOF (po-
sitive ion): m/z 457.17 [M+Na]+ (calcd 457.13). Anal. Calcd for
0
0
C16H27NaO12 (434.37): C, 44.24; H, 6.27. Found: C, 44.30; H, 6.29.
(158 mg, 78%). [a] +39.4 (c 0.13, MeOH); Rf 0.33 (85:15 CH2Cl2–
D
MeOH); 1H NMR (CD3OD, 360 MHz): d (ppm) 7.46–7.21 (m, 15H,
arom.), 4.51–3.60 (m, 18H), 3.25 (s, 3H, OCH3); 13C NMR (CD3OD,
90 MHz): d (ppm) 140.4, 139.6 139.5 (3 ꢃ Cq arom.), 129.4–128.4
(arom.), 104.2 (C-10), 99.0 (C-1), 81.4, 81.1, 78.1, 77.6, 76.4, 75.4,
73.5, 71.4 (skeleton carbons), 76.1, 74.4, 74.0 (3 ꢃ PhCH2), 69.9
(C-6), 55.5 (OCH3); MALDI-TOF (positive ion): m/z 685.39
[M+Na]+ (calcd 685.22). Anal. Calcd for C34H39NaO12 (662.66): C,
61.63; H, 5.93. Found: C, 61.54; H, 6.01.
3.7. Methyl (sodium 2,3,4-tri-O-methyl-b-
nate)-(1?4)-2,3,6-tri-O-sodium-sulfonato-a-D-glucopyranoside
(4)
D
-glucopyranosyluro
A solution of compound 19 (101 mg, 0.245 mmol) in DMF
(3 mL) was treated with the SO3ꢁpyridine complex (585 mg,
3.675 mmol) for 5 h at room temperature, then cold satd NaHCO3
solution was added in excess (pH >7). The resulting mixture was
concentrated in vacuo. Then MeOH (5 mL) was added and after
stirring for 20 min the insolubles were removed by filtration. The
filtrate was concentrated under reduced pressure. The crude prod-
uct was purified by Sephadex LH 20 column chromatography in
3.5. Methyl (sodium 2,3,4-tri-O-methyl-b-
nate)-(1?4)-2,3,6-tri-O-benzyl- -glucopyranoside (17) and
methyl (sodium 4-deoxy-2,3-di-O-methyl- -threo-hex-4-eno
-glucopyran
D-glucopyranosyluro
a
-D
a
-L
pyranosyluronate)-(1?4)-2,3,6-tri-O-benzyl-a-D
MeOH to give 4 (124 mg, 70%) as a white powder. [a] +37.9 (c
D
0.11, MeOH); Rf 0.34 (4:6 CH2Cl2–MeOH); 1H NMR (D2O,
500 MHz): d (ppm) 5.17 (d, 1H, J1,2 3.4 Hz, H-1), 4.67–4.63 (m,
2H, H-10, H-3), 4.39–4.37 (m, 3H, H-2, H-6a,b), 4.09 (m, 1H, H-5),
oside (18)
To a solution of compound 16 (117 mg, 0.183 mmol)) in DMF
(2 mL) at 0 °C were successively added 60% NaH (44 mg, 1.1 mmol)
and MeI (154 lL, 2.47 mmol). After 2 h stirring at this temperature,
4.03 (m, 1H, H-4), 3.70 (d, 1H, J4 5 10.0 Hz, H-50), 3.63, 3.61, 3.49,
3.48 (4 ꢃ s, 12H, 4 ꢃ OCH3), 3.45 (t, 1H, J 9.2 Hz, H-40), 3.88 (m,
1H, H-30), 3.23 (t, 1H, J 8.8 Hz H-20); 13C NMR (D2O, 125 MHz): d
(ppm) 175.7 (CO), 100.8 (C-10), 97.2 (C-1), 84.2 (C-30), 82.2 (C-20),
81.6 (C-4), 76.4 (C-50), 76.2 (C-3), 75.0 (C-2), 73.5 (C-4), 69.1 (C-
5), 66.1 (C-6), 61.1, 60.6, 56.0 (4 ꢃ OCH3); MALDI-TOF (positive
ion): m/z 763.14 [M+Na]+ (calcd 762.95). Anal. Calcd for
0
0
MeOH (0.5 mL) was added. The reaction mixture was stirred for
15 min, and the solvents were evaporated. The reaction gave a
mixture of 17 and 18 which were separated by silica gel chroma-
tography (1:1 n-hexane–acetone). Compound 17 (53 mg, 42%):
[a
]
D
+7.0 (c 0.13, CHCl3); Rf 0.59 (1:1 n-hexane–acetone); 1H
NMR (CDCl3, 360 MHz): d (ppm) 7.40–7.20 (m, 15H, arom.),
C16H24Na4O21S3 (740.50): C, 25.95; H, 3.27; S, 12.99. Found: C,
4.97–4.41 (m, 7H), 2.28 (m, 1H, J1 ,2 7.4 Hz, H-10), 3.85–3.65 (m,
5H), 3.53, 3.41, 3.31 (4 ꢃ s, 12H, 4 ꢃ OCH3), 3.56–3.33 (m, 2H),
3.01–2.80 (m, 3H); 13C NMR (CDCl3, 90 MHz): d (ppm) 138.3,
138.0, 137.7 (3 ꢃ Cq arom), 128.3–127.3 (arom.), 102.0 (C-10),
98.1 (C-1), 85.2, 83.2, 80.7, 79.8, 79.1, 76.4, 69.9 (skeleton carbons),
75.4, 73.4, 73.2 (3 ꢃ PhCH2), 67.8 (C-6), 60.4, 60.3, 60.0, 55.1
(4 ꢃ OCH3); MALDI-TOF (positive ion): m/z 727.19 [M+Na]+ (calcd
727.27). Anal. Calcd for C37H45NaO12 (704.74): C, 63.06; H, 6.44.
Found: C, 62.91; H, 6.33. Compound 17 was prepared also from
22 according to the method described for the synthesis of 16 in a
yield of 64%.
0
0
25.84; H, 3.26; S, 12.84.
3.8. Methyl (6-O-triphenylmethyl-b-D-glucopyranosyl)-(1?4)-
2,3,6-tri-O-benzyl-a-D-glucopyranoside (20)
To a solution of compound 15 (1.11 g, 1.77 mmol) in pyridine
(18 mL) triphenylmethyl chloride (988 mg, 3.54 mmol) was added.
The mixture was stirred for 24 h at room temperature. After com-
pletion of the reaction, the mixture was concentrated in vacuo.
The crude product was purified by silica gel chromatography
(75:25 CH2Cl2–acetone + 1% Et3N) to give 20 (1.27 g, 83%) as a col-
Compound 18 (50 mg, 40%): [
a] +42.8 (c 0.60, CHCl3); Rf 0.42
D
ourless syrup. [
a
]
D ꢂ8.4 (c 0.27, CHCl3); Rf 0.54 (75:25 CH2Cl2–ace-
(1:1 n-hexane–acetone); 1H NMR (CDCl3, 360 MHz): d (ppm)
tone); 1H NMR (CDCl3, 360 MHz): d (ppm) 8.54–7.25 (m, 30H,
arom.), 4.88–4.44 (m, 8H), 4.05–3.57 (m, 4H), 3.55–3.39 (m, 7H),
3.32 (s, 3H, OCH3), 3.29–3.21 (m, 4H); 13C NMR (CDCl3, 90 MHz):
d (ppm) 149.5–123.6 (arom.), 102.8 (C-10), 98.1 (C-1), 86.9 (Ph3C),
80.5, 79.4, 76.4, 76.1, 74.3, 73.9, 71.8, 69.4 (skeleton carbons),
74.7, 73.5, 73.3 (3 ꢃ PhCH2), 68.4, 63.8 (2 ꢃ C-6), 55.0 (OCH3);
MALDI-TOF (positive ion): m/z 891.35 calcd for [M+Na]+ (calcd
891.37). Anal. Calcd for C53H56O11 (869.01): C, 73.25; H, 6.50.
Found: C, 73.15; H, 6.38.
7.32–7.29 (15H, arom.), 6.07 (d, 1H J3 ,4 2.9 Hz, H-40), 4.9–4.85
(m, 3H), 4.81–4.42 (m, 5H), 3.94–3.81 (m, 4H), 3.78–3.51 (m,
3H), 3.45, 3.42, 3.37 (3 ꢃ s, 9H, 3 ꢃ OCH3), 3.21 (t, 1H, J 6.6 Hz);
13C NMR (CDCl3, 90 MHz): d (ppm) 161.9 (CO), 140.6 (C-50),
138.1, 138.0, 137.6 (3 ꢃ Cq arom.), 128.4–127.9 (arom.), 109.2 (C-
40), 101.3 (C-10), 98.2 (C-1), 79.6, 79.5, 79.0, 76.9, 76.6, 69.7 (skele-
ton carbons), 75.8, 73.5, 73.4 (3 ꢃ PhCH2), 67.8 (C-6), 59.9, 56.9,
55.3 (3 ꢃ OCH3); MALDI-TOF (positive ion): m/z 695.33 [M+Na]+
0
0

M. Herczeg et al. / Carbohydrate Research 346 (2011) 1827–1836
1833
3.9. Methyl (2,3,4-tri-O-methyl-6-O-triphenylmethyl-b-
D
-gluco
give 784 mg (78%) of 28 as a colourless syrup. [
a
]
+39.6 (c 0.27,
D
pyranosyl)-(1?4)-2,3,6-tri-O-benzyl- -glucopyranoside (21)
a-
D
CHCl3); Rf 0.30 (7:3 n-hexane–EtOAc); 1H NMR (CDCl3, 360 MHz):
d (ppm) 7.36–7.24 (m, 10H, arom.), 5.38, 5.32 (2 ꢃ s, 2H, CH2@),
4.77 (d, 1H, PhCH2), 4.71 (d, 1H, J1,2 3.6 Hz, H-1), 4.62–4.52 (m,
3H, PhCH2), 4.07 (d, 1H, J4,5 8.9 Hz, H-4), 3.94 (d, 1H, H-2), 3.74,
3.67 (2 ꢃ dd, 2H, H-6a,b), 3.58 (m, 1H, H-5), 3.38 (s, 3H, OCH3),
2.65 (s, 1H, OH); 13C NMR (CDCl3, 90 MHz): d (ppm) 143.0 (C-3),
137.8–127.6 (arom.), 105.3 (CH2@), 98.5 (C-1), 76.3 (C-2), 72.3
(C-4), 73.5, 71.9 (2 ꢃ PhCH2), 70.1 (C-6), 69.4 (C-5), 55.1 (OCH3);
Anal. Calcd for C22H26O5 (370.44): C, 71.33; H, 7.07. Found: C,
71.25; H, 7.05.
Compound 20 (1.20 g, 1.38 mmol) was methylated according to
the method described for the synthesis of 17. The crude product
was purified by silica gel chromatography (65:35 n-hexane–
EtOAc + 1% Et3N) to give 21 (1.08 g, 86%) as a colourless syrup.
[
a]
D
+9.1 (c 0.16, CHCl3); Rf 0.42 (65:35 n-hexane–EtOAc); 1H
NMR (CDCl3, 360 MHz): d (ppm) 7.50–7.09 (m, 30H, arom.),
5.03–4.47 (m, 8H), 4.34 (m, 1H), 4.01–3.92 (m, 2H), 3.78–3.70
(m, 4H), 3.59, 3.45, 3.39, 3.23 (4 ꢃ s, 12H, 4 ꢃ OCH3), 3.55–3.26
(m, 3H), 2.99–2.84 (m, 2H); 13C NMR (CDCl3, 90 MHz): d (ppm)
143.8–126.7 (arom.), 102.2 (C-10), 98.3 (C-1), 86.1 (Ph3C), 86.9,
84.4, 79.8, 79.5, 79.2, 75.6, 74.4, 70.0 (skeleton carbons), 74.6,
73.4, 73.2 (3 ꢃ PhCH2), 68.6, 61.4 (2 ꢃ C-6), 60.6, 60.4, 60.2, 55.2
(4 ꢃ OCH3); MALDI-TOF (positive ion): m/z 933.41 [M+Na]+ (calcd
933.42). Anal. Calcd for C56H62O11 (911.08): C, 73.82; H, 6.86.
Found: C, 73.69; H, 6.78.
3.13. Methyl 2,6-di-O-benzyl-3-deoxy-3-C-(triethylammonium
sulfonatomethyl)-a-D-glucopyranoside (27)
To a solution of compound 28 (500 mg, 1.35 mmol) in aqueous
EtOH (70%, 80 mL) NaHSO3 (1.4 g, 13.5 mmol) and tert-butyl per-
oxybenzoate (128 lL, 0.675 mmol) were added and the mixture
was heated under reflux for 4 h. After cooling, triethylamine was
added, the mixture was diluted with EtOH, the insolubles were re-
moved by filtration and the filtrate was concentrated in vacuo. The
crude product was purified by silica gel chromatography (90:9:1
CH2Cl2–MeOH–Et3N) to give 553 mg (74%) of 279 as a colourless
3.10. Methyl (2,3,4-tri-O-methyl-b-
D-glucopyranosyl)-(1?4)-
2,3,6-tri-O-benzyl- -glucopyranoside (22)
a-D
A solution of compound 21 (1.00 g, 1.01 mmol) in AcOH (80%,
18 mL) was stirred for 2 h at 50 °C. The mixture was then concen-
trated in vacuo. The crude product was purified by silica gel chro-
matography (9:1 CH2Cl2–acetone) to give 22 (716 mg, 98%) as a
syrup. [
a]
+50.1 (c 0.22, CHCl3), lit.9 +50.3. The 1H and 13C NMR
D
data were in agreement with those published.9
colourless syrup. [
a
]
+21.3 (c 0.15, CHCl3); Rf 0.42 (9:1 CH2Cl2–
3.14. Methyl (b-D-glucopyranosyl)-(1?4)-2,3-di-O-benzyl-6-
D
acetone); 1H NMR (CDCl3, 360 MHz): d (ppm) 7.37–7.24 (m, 15H
deoxy-6-C-(triethylammonium sulfonatomethyl)-a-D-gluco
arom.), 4.97–4.45 (m, 7H), 4.23 (d, 1H, J1 2 7.5 Hz, H-10), 3.91–
3.67 (m, 6H), 3.59, 3.50, 3.46, 3.38 (4 ꢃ s, 12H, 4 ꢃ OCH3), 3.58–
3.26 (m, 3H), 2.98–2.95 (m, 4H); 13C NMR (CDCl3, 90 MHz): d
(ppm) 139.2–126.9 (arom.), 102.4 (C-10), 98.2 (C-1), 86.3, 84.1,
79.9, 79.6, 78.8, 77.1, 74.6, 70.0 (skeleton carbons), 75.0, 73.4,
73.2 (3 ꢃ PhCH2), 67.8 (C-6), 61.7 (C-60), 60.5, 60.4, 60.1, 55.2
(4 ꢃ OCH3); MALDI-TOF (positive ion): m/z 691.28 [M+Na]+ (calcd
691.31). Anal. Calcd for C37H48O11 (668.77): C, 66.45; H, 7.23.
Found: C, 66.36; H, 7.18.
pyranoside (33)9
0
0
Monitoring the proton deuterium exchange the initial proton
spectrum was recorded in DMSO, then D2O was added. 1H NMR
(DMSO, 400 MHz): d (ppm) 7.39–7.29 (m, 10H, arom.), 5.08 (d,
1H, J 4.5 Hz, 20-OH; 0.4H intensity 16 scans after adding D2O),
4.94–4.89 (m, 3H, 30-OH, 40-OH, PhCH2; 1.6H intensity 16 scans
after adding D2O), 4.77 (d, 1H, J1,2 3.1 Hz, H-1), 4.71 (d, 1H, PhCH2),
0
0
0
4.61 (s, 2H, PhCH2), 4.48 (d, 1H, J1 ,2 7.5 Hz, H-1 ), 4.41–4.38 (m, 1H,
60-OH, the signal disappeared 16 scans after adding D2O), 3.72 (t,
1H, H-4), 3.61–3.46 (m, 3H), 3.43–3.35 (m, 4H), 3.29 (s, 3H,
OCH3), 3.18–3.15 (m, 1H, H-2), 3.08–3.03 (m, 6H, Et3N–CH2),
3.01–21.98 (m, 1H, H-20), 2.68–2.52 (m, 3H), 2.32 (m, 1H, H-6a),
1.77 (m, 1H, H-6b), 1.19 (m, 9H, Et3N–CH3); 13C NMR (D2O,
90 MHz): d (ppm) 138.9, 138.6 (Cq arom.), 129.5–128.9 (arom.),
103.2 (C-10), 98.2 (C-1), 80.8, 80.3, 80.0, 77.5, 76.9, 75.3, 71.2,
69.9 (skeleton carbons), 76.6, 74.0 (2 ꢃ PhCH2), 62.3 (C-60), 55.8
(OCH3), 48.3 (C-7), 47.6 (NCH2), 27.3 (C-6), 9.2 (NCH3).
3.11. 1,5-Anhydro-3-O-benzyl-4,6-O-benzylidene-2-deoxy-2-C-
methyl-D-arabino-hex-1-enitol (25)
To a solution of compound 23 (366 mg, 1 mmol) in CH2Cl2
(10 mL) at 0 °C Et3SiH (1.91 mL, 12.0 mmol) and BF3ꢁEt2O
(0.25 mL, 2.0 mmol) were added. The reaction mixture was stirred
at room temperature for 4 h. The mixture was diluted with CH2Cl2
(200 mL), washed with satd NaHCO3 (2 ꢃ 100 mL), dried and con-
centrated. The crude product was purified by silica gel chromatog-
raphy (4:1 n-hexane–EtOAc) to give 140 mg (41%) of 25 as white
3.15. Methyl 1,2,4-tri-O-acetyl-3-O-methyl-a,b-L-idopyranosy
needles. Mp: 110–112 °C; [
a
]
ꢂ1.0 (c 0.19, CHCl3); Rf 0.71 (97:3
luronate (37)
D
CH2Cl2–EtOAc); 1H NMR (CDCl3, 200 MHz): d (ppm) 7.54–7.20
(m, 10H, arom.), 6.17 (s, 1H, H-1), 5.62 (s, 1H, CHPh), 4.94–4.69
(dd, 2H, J 11.8 Hz, PhCH2), 4.36 (m, 1H, H6a), 4.24 (d, 1H, J3,4
7.3 Hz, H-3), 4.06 (dd, 1H, J4,5 9.4 Hz, H-4), 3.86–3.80 (m, 2H, H-
5, H-6b), 1.63 (s, 3H, 2-CH3); 13C NMR (CDCl3, 50 MHz): d (ppm)
A solution of 36 (5.0 g, 19.065 mmol) in aqueous 90% trifluoro-
acetic acid (40 mL) was kept at room temperature for 15 min,
evaporated to dryness, and the residue was coevaporated with
H2O (2 ꢃ 10 mL). The obtained crude product was acetylated using
pyridine (28 mL) and acetic anhydride (14 mL). After 24 h at room
temperature, the mixture was concentrated under reduced
pressure, and traces of pyridine and acetic anhydride were
coevaporated three times with toluene. Column chromatography
139.9 (C-1), 138.7, 137.4 (Cquat, arom), 128.9–125.9 (arom.),
110.3 (C-2), 101.1 (CHPh), 81.1 (C-4), 75.9 (C-3), 68.6 (C-5), 73.3
(PhCH2), 68.5 (C-6), 13.8 (2-CH3); MALDI-TOF (positive ion): m/z
361.42 [M+Na]+ (calcd 361.14). Anal. Calcd for C21H22O4 (338.41):
C, 74.54; H, 6.55. Found: C, 74.35; H, 6.46.
(1:1 n-hexane–EtOAc) gave 59 (2.39 g, 36% for 2 steps,
ꢀ1:10) as a colourless oil.
a:b
3.12. Methyl 2,6-di-O-benzyl-3-deoxy-3-C-methylene-
a-
D-ribo-
a
: Rf 0.38 (1:1 n-hexane–EtOAc); 1H NMR (CDCl3, 360 MHz): d
hexopyranoside (28)
(ppm) 6.22 (s, 1H, H-1), 5.18 (s, 1H), 4.89 (s, 1H), 4.87 (s, 1H),
3.80 (s, 3H, COOCH3), 3.63 (s, 1H, H-3), 3.54 (s, 3H, OCH3), 2.12,
2.09, 2.08 (3s, 9H, 3 ꢃ CH3); 13C NMR (CDCl3, 90 MHz): d (ppm)
169.6, 169.2, 168.4, 168.1 (4 ꢃ CO), 91.0 (C-1), 73.4, 67.5, 67.0,
65.3 (skeleton carbons), 58.4 (OCH3), 52.5 (COOCH3), 20.8, 20.7,
Compound 26 (1.0 g, 2.714 mmol) was converted to 28 by the
method described for the synthesis of 25. The crude product was
purified by silica gel chromatography (7:3 n-hexane–EtOAc) to

1834
M. Herczeg et al. / Carbohydrate Research 346 (2011) 1827–1836
20.6 (3 ꢃ CH3). Anal. Calcd for C14H20O10 (348.30): C, 48.28; H,
100.1, 97.7 (2 ꢃ C-1), 79.9, 79.6, 77.5, 74.6, 69.8, 68.3, 67.5, 66.8
(skeleton carbons), 74.7, 73.3, 73.2 (3 ꢃ PhCH2), 68.5 (C-6), 57.8,
54.9, 51.7 (3 ꢃ OCH3); MALDI-TOF (positive ion): m/z 691.35
[M+Na]+ (calcd 691.27). Anal. Calcd for C36H44O12 (668.73): C,
64.66; H, 6.63. Found: C, 64.59; H, 6.61.
5.79. Found: C, 48.19; H, 5.74.
b: Rf 0.30 (1:1 n-hexane–EtOAc); [a]D +22.3 (c 0.10, CHCl3); mp
123–124 °C. The 1H and 13C NMR data have been published.9 Anal.
Calcd for C14H20O10 (348.30): C, 48.28; H, 5.79. Found: C, 48.37; H,
5.77.
3.19. Methyl (methyl 2,3,4-tri-O-methyl-a-L-idopyranosyluro
3.16. Methyl 2,4-di-O-acetyl-3-O-methyl-
a,b-
L-idopyranosyluro
nate)-(1?4)-2,3,6-tri-O-benzyl- -glucopyranoside (42)
a-D
nate (38)
To a solution of 41 (134 mg, 0.200 mmol) in DMF (4.5 mL) at rt
To a solution of 37 (2.0 g, 5.742 mmol) in THF (50 mL) benzyl
amine (2.51 mL, 22.97 mmol) was added. The reaction mixture
was stirred for 4 h and monitored by TLC. The reaction was poured
into 1 M aq HCl (50 mL), and the aqueous phase was extracted with
EtOAc (3 ꢃ 100 mL). The combined organic phases were dried, and
evaporated to dryness. Column chromatography (85:15 CH2Cl2–
were successively added freshly prepared Ag2O (370 mg, 1.6 mmol,
4 equiv/OH) and MeI (37 lL, 0.6 mmol, 1.5 equiv/OH). After 96 h of
stirring at this temperature, the mixture was diluted with CH2Cl2,
filtered off through a pad of Celite and the filtrate was concentrated
under reduced pressure. The crude product was purified by silica
gel chromatography (95:5 CH2Cl2–acetone) to give 42 as a colour-
acetone) gave 38 (1.39 g, 79%) as a colourless oil. [
a
]
+4.7 (c
less syrup (105 mg, 76%). [
a
]
ꢂ19.6 (c 0.10, CHCl3); Rf 0.49 (95:5
D
D
0.10, CHCl3); Rf 0.43 (85:15 CH2Cl2–acetone); 1H NMR (CDCl3,
500 MHz): d (ppm) 5.33 (s, 0.6H), 5.31 (s, 0.4H), 5.19 (s, 0.6H),
5.12 (s, 0.4H), 5.10 (s, 0.6H), 4.99 (s, 0.6H), 4.88 (s, 0.4H), 4.82 (s,
0.6H), 4.62 (s, 0.8H), 3.78, 3.77, 3.60, 3.55 (4 ꢃ s, 6H, 4 ꢃ OCH3),
3.45, 3.44 (2 ꢃ s, 1H), 2.08, 2.06, 2.04 (3 ꢃ s, 6H, 4 ꢃ CH3); 13C
CH2Cl2–acetone); 1H NMR (CDCl3, 360 MHz): d (ppm) 7.39–7.27
(m, 15H, arom.), 5.17 (d, 1H, J 5.3 Hz), 4.87 (dd, 2H, PhCH2),
4.75–4.55 (m, 6H), 3.91–3.56 (m, 7H), 3.53, 3.48, 3.42, 3.41, 3.36
(5 ꢃ s, 15H, 5 ꢃ OCH3), 3.35–3.30 (m, 1H), 3.04–2.98 (m, 1H); 13C
NMR (CDCl3, 90 MHz): d (ppm) 169.5 (CO), 139.0, 138.1, 137.9
(3 ꢃ Cq arom.), 128.2–127.0 (arom.), 99.6, 98.0 (2 ꢃ C-1), 81.3,
79.9, 79.6, 79.5, 78.8, 75.8, 70.7, 70.1 (skeleton carbons), 75.1,
73.4, 73.3 (3 ꢃ PhCH2), 68.2 (C-6), 59.7, 59.5, 58.8, 55.1, 51.5
(5 ꢃ OCH3); MALDI-TOF (positive ion): m/z 719.35 [M+Na]+ (calcd
719.30). Anal. Calcd for C38H48O12 (696.78): C, 65.50; H, 6.94.
Found: C, 65.64; H, 6.97.
NMR (CDCl3, 125 MHz): d (ppm) 92.6, 91.7 (C-1a, C-1b), 74.8,
74.2, 72.2, 67.4, 66.6, 66.5, 65.3 (skeleton carbons), 58.9, 58.6
(2 ꢃ OCH3), 52.4, 52.3 (2 ꢃ COOCH3), 20.6, 20.5, 20.4, 20.3
(4 ꢃ CH3); Anal. Calcd for C12H18O9 (306.27): C, 47.06; H, 5.92.
Found: C, 46.93; H, 5.98.
3.17. Methyl (methyl 2,4-di-O-acetyl-3-O-methyl-
a
-L
-idopyr
anosyluronate)-(1?4)-2,3,6-tri-O-benzyl-
a-D
-glucopyranoside
3.20. Methyl (sodium 2,3,4-tri-O-methyl-a-L-idopyranosyluro
(40)
nate)-(1?4)-(2,3,6-tri-O-benzyl- -glucopyranoside (43)
a-D
A mixture of compound 13 (400 mg, 0.861 mmol), imidate 39
(582 mg, 1.292 mmol) and 4 Å molecular sieves in dry CH2Cl2
(8 mL) was stirred for 15 min at room temperature, then cooled
to ꢂ20 °C under argon. TMSOTf (0.1 M in CH2Cl2, 1.29 mL,
0.13 mmol) was added, and the reaction mixture was allowed to
warm up to 0 °C in 30 min. After completion of the reaction it
was quenched by addition of Et3N (0.2 mL). The reaction mixture
was then filtered and concentrated. The crude product was purified
by silica gel chromatography (7:3 n-hexane–acetone) to give 40 as
Compound 42 (184 mg, 0.260 mmol) was dissolved in MeOH
(20 mL) and treated with 0.1 M aq NaOH solution (20 mL). After
24 h stirring at rt the TLC showed complete conversion of carbox-
ylic ester into sodium salt. The mixture was neutralized with acetic
acid, concentrated in vacuo and the residue was purified by silica
gel chromatography (95:5 CH2Cl2–MeOH) to give 43 as a colourless
syrup (126 mg, 68%), [
a]
+23.1 (c 0.19, MeOH); Rf 0.25 (9:1
D
CH2Cl2–MeOH); 1H NMR (CD3OD, 360 MHz): d (ppm) 7.34–7.25
(m, 15H, arom.), 5.17 (d, 1H, J 4.0 Hz), 4.88 (s, 1H), 4.73–4.50 (m,
7H), 3.81–3.70 (m, 5H), 3.58–3.53 (m, 2H), 3.47–3.44 (m, 1H),
3.48, 3.39, 3.36, 3.32 (4 ꢃ s, 12H, 4 ꢃ OCH3), 3.15–3.11 (m, 1H);
13C NMR (CD3OD, 90 MHz): d (ppm) 174.0 (CO), 140.1, 139.5,
139.4 (3 ꢃ Cq arom.), 129.3–128.3 (arom.), 100.7, 98.8 (2 ꢃ C-1),
81.3, 80.9, 81.0, 80.9, 79.7, 79.1, 77.1, 71.7 (skeleton carbons),
76.2, 74.4, 73.9 (3 ꢃ PhCH2), 69.8 (C-6), 59.7, 59.3, 59.0, 55.5
(4 ꢃ OCH3); MALDI-TOF (positive ion): m/z 705.42 (uronic acid)
[M+Na]+ (calcd 705.29). Anal. Calcd for C37H45NaO12 (704.74): C,
63.06; H, 6.44. Found: C, 63.07; H, 6.40.
a colourless oil (609 mg, 94%). [
a]
ꢂ25.1 (c 0.15, CHCl3); Rf 0.48
D
(1:1 n-hexane–EtOAc); 1H NMR (CDCl3, 360 MHz): d (ppm) 7.35–
7.26 (m, 15H, arom.), 5.11–4.98 (m, 4H), 4.79–4.49 (m, 7H),
3.99–3.71 (m, 6H), 3.63–3.58 (m, 1H), 3.52, 3.36, 3.33 (3 ꢃ s, 9H,
3 ꢃ OCH3), 2.00, 1.99 (2 ꢃ s, 6H, 2 ꢃ CH3); 13C NMR (CDCl3,
90 MHz): d (ppm) 169.8, 169.5, 168.5 (3 ꢃ CO), 138.6, 137.9,
137.8 (3 ꢃ Cq arom.), 128.2–126.9 (arom.), 97.8, 96.8 (2 ꢃ C-1),
80.0, 79.5, 74.2, 73.9, 69.9, 67.1, 66.7, 65.8 (skeleton carbons),
74.8, 73.2, 73.0 (3 ꢃ PhCH2), 68.0 (C-6), 58.2, 55.0, 51.8 (3 ꢃ OCH3),
20,7, 20.6 (2 ꢃ CH3); MALDI-TOF (positive ion): m/z 775.39
[M+Na]+ (Calcd 775.29). Anal. Calcd for C40H48O14 (752.80): C,
63.82; H, 6.43. Found: C, 63.68; H, 6.46.
3.21. Methyl (sodium 2,3,4-tri-O-methyl-a-L-idopyranosyluro
nate)-(1?4)- -glucopyranoside (44)
a-D
3.18. Methyl (methyl 3-O-methyl-
a
-
L
-idopyranosyluronate)-
Compound 43 (110 mg, 0.156 mmol) was debenzylated accord-
ing to the method described for the synthesis of 19. The crude prod-
uct was purified by silica gel chromatography (1:1 CH2Cl2–MeOH) to
(1?4)-2,3,6-tri-O-benzyl- -glucopyranoside (41)
a-D
Compound 40 (88 mg, 0.117 mmol) was deacetylated as de-
scribed for the synthesis of 15. The crude product was purified
by silica gel chromatography (85:15 CH2Cl2–acetone) to give 41
give 44 as a white powder (58 mg, 85%). [a]D +54.0 (c 0.10, MeOH); Rf
0.21 (1:1 CH2Cl2–MeOH); 1H NMR (D2O, 360 MHz): d (ppm) 5.06 (s,
1H), 4.80 (s, 1H), 4.65 (s, 1H), 3.88–3.85 (m, 1H), 3.81–3.56 (m, 8H),
3.52, 3.49, 3.42, 3.40 (4 ꢃ s, 12H, 4 ꢃ OCH3); 13C NMR (D2O,
90 MHz): d (ppm) 175.9 (CO), 99.3, 99.0 (2 ꢃ C-1), 77.4, 77.2, 76.9,
75.9, 71.7, 71.4, 70.5, 69.6 (skeleton carbons), 60.1 (C-6), 58.3,
57.9, 54.9 (4 ꢃ OCH3); MALDI-TOF (positive ion): m/z 457.32 (so-
dium salt) [M+Na]+ (calcd 457.13). Anal. Calcd for C16H27NaO12
(434.37): C, 44.24; H, 6.27. Found: C, 44.31; H, 6.30.
as a colourless oil (74 mg, 95%). [
a]
ꢂ17.3 (c 0.15, CHCl3); Rf
D
0.50 (85:15 CH2Cl2–acetone); 1H NMR (CDCl3, 360 MHz): d (ppm)
7.33–7.25 (m, 15H, arom.), 5.03 (s, 1H), 4.93–4.51 (m, 8H), 3.92–
3.71 (m, 5H), 3.68–3.54 (m, 5H), 3.47–3.44 (m, 1H), 3.40, 3.33
(2 ꢃ s, 9H, 3 ꢃ OCH3), 13C NMR (CDCl3, 90 MHz): d (ppm) 170.5
(CO), 138.7, 137.8. 137.5 (3 ꢃ Cq arom.), 128.2–126.8 (arom.),

M. Herczeg et al. / Carbohydrate Research 346 (2011) 1827–1836
1835
644–649; (c) Thunberg, L.; Backström, G.; Lindahl, U. Carbohydr. Res. 1982, 100,
393–410.
3.22. Methyl (sodium 2,3,4-tri-O-methyl-
nate)-(1?4)-2,3,6-tri-O-(sodium sulfonato)-a-D
side (8)
a
-
L
-idopyranosyluro
-glucopyrano
3. (a) Sina, P.; Jacquinet, J.-C.; Petitou, M.; Duchaussoy, P.; Lederman, I.; Choay, J.;
Torri, G. Carbohydr. Res. 1984, 132, C5–C9; (b) Petitou, M.; Duchaussoy, P.;
Lederman, I.; Choay, J.; Sina, P.; Jacquinet, J.-C.; Torri, G. Carbohydr. Res. 1986,
147, 221–236; (c) van Boeckel, C. A. A.; Petitou, M. Angew. Chem., Int. Ed. Engl.
1993, 32, 1671–1690.
4. (a) van Boeckel, C. A. A.; Petitou, M. Angew. Chem., Int. Ed. Engl. 2004, 43, 3118–
3133; (b) Cheng, J. M. W. Clin. Ther. 2002, 24, 1757–1769.
5. Westerduin, P.; van Boeckel, C. A. A.; Basten, J. E. M.; Broekhoven, M. A.; Lucas,
H.; Rood, A.; van der Heiden, H.; van Amsterdam, R. G. M.; van Dinther, T. G.;
Meuleman, D. G.; Visser, A.; Vogel, G. M. T.; Damm, J. B. L.; Overklift, G. T.
Bioorg. Med. Chem. 1994, 2, 1267–1280.
Compound 44 (50 mg, 0.115 mmol) was O-sulfated according to
the method described for the synthesis of 4. The crude product was
purified by Sephadex LH-20 column chromatography eluted with
H2O to give 8 as a white powder (80 mg, 94%). [
a] +36.8 (c 0.15,
D
MeOH); Rf 0.10 (8:5:1 CH2Cl2–MeOH-H2O); 1H NMR (D2O,
0
0
500 MHz): d (ppm) 5.19 (d,1H, J1,2 3.5 Hz, H-1), 5.06 (d, 1H, J1 ,2
3.3 Hz, H-10), 4.82 (d, 1H, J4 ,5 2.7 Hz, H-50), 4.67 (t, 1H, J 9.4 Hz,
H-3), 4.43 (m, 1H, H-6a), 4.41 (m, 1H, H-2), 4.32 (m, 1H, H-6b),
4.12 (m, 1H, H-5), 4.03 (t, 1H, J 9.7 Hz, H-4), 3.82 (m, 1H, H-40),
3.67 (m, 1H, H-30), 3.58, 3.57, 3.51, 3.48 (4 ꢃ s, 12H, 4 ꢃ OCH3),
6. Chen, Ch.; Yu, B. Bioorg. Med. Chem. Lett. 2009, 19, 3875–3879.
7. (a) Gómez-Outes, A.; Lecumberri, R.; Pozo, C.; Rocha, E. Curr. Vasc. Pharmacol.
2009, 7, 309–329; (b) Harenberg, J. Expert Rev. Clin. Pharmacol. 2010, 3, 9–16.
8. Savi, P.; Herault, J. P.; Duchaussoy, P.; Millet, L.; Schaeffer, P.; Petitout, M.; Bono,
F.; Herbert, J. M. J. Thromb. Haemost. 2008, 6, 1697–1706.
9. Herczeg, M.; Lázár, L.; Borbás, A.; Lipták, A.; Antus, S. Org. Lett. 2009, 11, 2619–
2622.
10. Lázár, L.; Herczeg, M.; Fekete, A.; Borbás, A.; Lipták, A.; Antus, S. Tetrahedron
Lett. 2010, 51, 6711–6714.
11. Lemieux, R. U. In Methods in Carbohydrate Chemistry; Whistler, R. L., Wolfrom,
M. L., BeMiller, J. N., Eds.; Academic Press, 1963; Vol. II, pp 221–222.
12. Ek, M.; Garegg, P. J.; Hultberg, H.; Oscarson, S. J. Carbohydr. Chem. 1983, 2, 331–
336.
13. (a) Davis, N. J.; Flitsch, S. L. Tetrahedron Lett. 1993, 34, 1181–1184; (b) de Noy,
A. E. J.; Besemer, A. C.; van Bekkum, H. Carbohydr. Res. 1995, 269, 89–98.
14. Borbás, A.; Csávás, M.; Szilágyi, L.; Májer, G.; Lipták, A. J. Carbohydr. Chem. 2004,
23, 133–146.
15. Wenz, G.; Höfler, T. Carbohydr. Res. 1999, 322, 153–165.
16. Sarda, P.; Olesker, A.; Lukács, G. Carbohydr. Res. 1992, 229, 161–165.
17. Yoshimura, J.; Kawauchi, N.; Yasumori, T.; Sato, K.; Hashimoto, H. Carbohydr.
Res. 1984, 133, 255–274.
0
0
13
3.50 (m, 1H, H-2); C NMR (D2O, 125 MHz) d (ppm) 99.6 (C-10),
97–2 (C-1), 79.0 (C-40), 78.1 (C-30), 78.0 (C-20), 76.1 (C-3), 75.3
(C-2), 73.3 (C-4), 70.9 (C-50), 68.8 (C-5), 66.3 (C-6), 58.8, 58.4,
58.0, 55.3 (4 ꢃ OCH3); MALDI-TOF (positive ion): m/z 763.03 (tet-
rasodium salt) [M+Na]+ (calcd 762.95). Anal. Calcd for
C
16H24Na4O21S3 (740.50): C, 25.95; H, 3.27; S, 12.99. Found: C,
25.76; H, 3.22; S, 12.89.
3.23. Methyl (methyl 2,3,4-tri-O-methyl-a-L-idopyranosyluro
nate)-(1?4)-3,6-di-O-benzyl-2-deoxy-2-C-(sodium sulfonatom
ethyl)- -glucopyranoside (47); and methyl (methyl 4-deoxy-
2,3-di-O-methyl- -threo-hex-4-enopyranosyluronate)-(1?4)-
a
-D
a-L
18. Debenham, S. D.; Toone, E. J. Tetrahedron: Asymmetry 2000, 11, 385–387.
3,6-di-O-benzyl-2-deoxy-2-C-(sodium sulfonatomethyl)-a-D-
ˇ
19. (a) Lipták, A.; Neszmélyi, A.; Kovác, P.; Hirsch, J. Tetrahedron 1981, 37, 2379–
glucopyranoside (48)
2382; (b) Gigg, R.; Conant, R. J. Carbohydr. Chem. 1982–1983, 1, 331–336; (c)
Gigg, R.; Conant, R. Carbohydr. Res. 1982, 104, C14–C17.
20. Jakab, Zs.; Mándi, A.; Borbás, A.; Bényei, A.; Komáromi, I.; Lázár, L.; Antus, S.;
Lipták, A. Carbohydr. Res. 2009, 344, 2444–2453.
21. van den Bos, L. J.; Codée, J. D. C.; Litjens, R. E. J. N.; Dinkelaar, J.; Overkleeft, H.
S.; van der Marel, G. A. Eur. J. Org. Chem. 2007, 3963–3976.
Compound 46 (190 mg, 0.28 mmol) was methylated according
to the method described for the synthesis of 17 to give a mixture
of 47 and 48 (140 mg) which could not be separated by silica gel
chromatography. Rf of the mixture: 0.71 (7:3 CH2Cl2–MeOH),
0.22 (n-hexane–EtOAc).
22. For recent syntheses of L-iduronic acid or L-idose building blocks, see: (a)
Tabeur, C.; Machetto, F.; Mallet, J.-M.; Duchaussoy, P.; Petitou, M.; Sinay, P.
Carbohydr. Res. 1996, 281, 253–275; (b) Adinolfi, A.; Barone, G.; DeLorenzo, F.;
Iadonisi, A. Synlett 1999, 1316–1318; (c) Lubineau, A.; Gavard, O.; Alais, J.;
Bonnaffé, D. Tetrahedron Lett. 2000, 41, 307–311; (d) Hung, S.-C.; Thopate, S. R.;
Chi, F.-C.; Chang, S.-W.; Lee, J.-C.; Wang, C.-C.; Wen, Y.-S. J. Am. Chem. Soc. 2001,
123, 3153–3154; (e) Lohman, G. J. S.; Hunt, D. K.; Högermaier, J. A.; Seeberger,
P. H. J. Org. Chem. 2003, 68, 7559–7561; (f) Ke, W.; Whitfield, D. M.; Gill, M.;
Laroque, S.; Yu, S.-H. Tetrahedron Lett. 2003, 44, 7767–7770; (g) Gavard, O.;
Hersant, Y.; Alais, J.; Duverger, V.; Dilhas, A.; Bascou, A.; Bonnaffé, D. Eur. J. Org.
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Compound 47: 13C NMR (CDCl3, 125 MHz): d (ppm) 169.9 (CO),
138.3–127. (arom.), 99.8, 98.7 (C-1, C-10), 82.2, 80.3, 79.2, 77.4,
76.5, 71.2, 70.6 (skeleton carbons), 73.2 (2 ꢃ PhCH2) 68.3 (C-6),
60.0, 59.7, 58.8, 55.2, 51.8 (5 ꢃ OCH3), 48.1 (CH2SO3ꢂ), 42.4 (C-
2);
MALDI-TOF
(positive
ion):
m/z
729.31
[M+Na]+
(calcd 729.25).
For NMR measurments 35 mg of the mixture of 47 and 48 in
DMF (1 mL) was treated with NaH (5 mg) to give 48 as a colourless
oil; 1H NMR (CDCl3, 500 MHz); d (ppm) 7.32–7.15 (m, 10H, arom.),
5.95 (d, 1H, J3 ,4 3.0 Hz, H-40), 5.17 (s, 1H), 4.90–4.41 (m, 5H), 4.07
(t, 1H, J 9.5 Hz), 3.81–3.57 (m, 4H), 3.48, 3.44, 3.41, 3.20 (4 ꢃ s,
12H, 4 ꢃ OCH3), 3.23–3.20 (m, 1H), 3.06–2.90 (m, 3H), 2.53 (m,
1H, H-2); 13C NMR (CDCl3, 125 MHz): d (ppm) 162.5 (CO), 141.8
(C-50), 138.1, 137.9 (2 ꢃ Cq arom.), 128.1–127.6 (arom.), 109.0 (C-
40), 101.4, 99.0 (C-1, C-10), 79.3, 78.6, 76.6, 70.4 (skeleton carbons),
73.2 (2 ꢃ PhCH2), 68.1 (C-6), 59.9, 57.0, 55.3, 52.5 (4 ꢃ OCH3), 47.7
(CH2SO3ꢂ), 42.3 (C-2); MALDI-TOF (positive ion): m/z 697.31
[M+Na]+ (calcd 697.21).
0
0
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Acknowledgements
The work is supported by the TÁMOP 4.2.1/B-09/1/KONV-2010-
0007 project. The project is co-financed by the European Union and
the European Social Fund. Financial support of the Hungarian Re-
search Fund (K 62802) is also acknowledged.
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